Heparanase, Academia, AB733

In the study reported in Spyrou et al., Mol Cancer Ther, 2017 Aug; 16(8): 1705-1716, this antibody was used for detection of heparanse in

1)Western blot of lysates of human CNS embryonal tumors, medulloblastomas, control embryonal brain tissue and tumor cell lines D283, D324 and PFSK-1.
dilution 1:500 in 1x TBST with 5% w/v nonfat dry milk
2) IHC of human CNS embryonal tumors, medulloblastomas and control brain tissue
dilution 1:200 in blocking/permeabilization buffer containing 5% normal goat serum (NGS) and 0.1-0.3% Triton-X 100 in 1xPBS
3) immunofluorescent staining of fixed PFA (4%) fixed cells (tumor cell lines D283, D324 and PFSK-1)
dilution 1:200 in blocking/permeabilization buffer containing 5% normal goat serum (NGS) and 0.1-0.3% Triton-X 100 in 1xPBS)

The antibody was provided by Dr. Robert Heinrikson (Pfizer Inc., Kalamazoo), and first described in Fairbanks et al. October 15, 1999 The Journal of Biological Chemistry 274, 29587-29590.

Based on the experimental part done in Vlodavsky’s Laboratory, the anti-HPSE ab733 was raised against a 14 amino acid peptide mapped at the N-terminus region of the 50 kDa heparanase (Lys158-Asn171). This antibody preferentially recognizes the active 50 kDa heparanase form by means of immunoblotting, immunostaining, immunoprecipitation, and labels heparanase in archive paraffin sections subjected to immunohistochemistry.